Snake bite, a major socio-medical problem
of south east asian countries is still depending on the usage of antisera as
the one and only source of treatment, which has its own limitations.In India, mostly in rural areas,
health centres are inadequate and the snake bite victims mostly depend on
traditional healers and herbal antidotes, as an alternative treatment. The
present review has been focussed on the varied folk and traditional herbs and
their antisnake venom compounds, which might be a stepping stone in
establishing the future therapy against snake bite treatment and management.

Glycoprotein T 11 target structure (T11TS),
derived from sheep erythrocyte membrane, directly interacts with T cells to
activate them to enter in the brain. When untreated, glioma exerts an
immune-suppressive environment in its vicinity by secreting prostaglandin E2
(PGE2), IL-10, tumor growth factor β, gangliosides etc. to
dampen the immune attack. But exogenous administration of T11TS reverses the situation
to pro-inflammatory immune active state by expressing enhanced IL-12 and tumor
necrosis factor α (TNF-α) production and suppression of IL-4 and
IL-10 levels. The T11TS activated lymphocytic accumulation along the capillary
endothelium in brain and their penetration in the matrix was evident from
histological sections. IL-6 with TNF-α facilitates leukocyte migration to
glioma site to exert cytotoxic effector function. Brain infiltrated lymphocytes
offer cytotoxic proximity to neoplastic glial cells, which lead them to
apoptosis. In the Th1 dominated microenvironment microglial cells was found
with enhanced phagocytic functions. Initially infiltrated lymphocytes with
microglia showed increased production of TNF-α, interferon γ
(IFN-γ) to facilitate their effector actions. Repeated dosing of T11TS
shows glioma abrogation in rat model, but also a resurgence of
anti-inflammatory cytokine environment found with increased IL-4, IL-10 and
decreased IL-12, IL-6, TNF-α. This is a unique homeostatic regulation of
total immune system after T11TS mediated carnage of glioma. The resultant
balance of cytokines between interacting glioma cells, T cells and microglia in
T11TS induced condition determines the success of its immunotherapeutic effect
in glioma.

1,3Department
of Microbiology, 2Department of Animal Biotechnology, G. B. Pant University of Agriculture and Technology,
College of Veterinary Sciences Pantnagar 263 145, India

Received 17 March 2010; revised 13 May 2010

The R2B strain of virus of new castle disease virus
(NDV) was propagated in 9-11 day old embryonated chicken eggs via allantoic
cavity route and after seven serial passages virus was purified from allantoic
fluid.Purified virus was analyzed by sodium dodecyl sulfate polyacrylamide
gel electrophoresis which yielded six major polypeptides ranging from 38-200
kDa. Protein fractions, corresponding to 75 and 56kDa, resembling
haemagglutinin-neuraminidase (HN) and fusion (F) proteins were used to
ascertain their immunization potential. Immunization of viral proteins was
compared with the whole virus vaccine. Among different group of birds, highest
haemagglutination inhibition (HI) and enzyme
linked immunosorbent assay (ELISA) titers were obtained in birds
immunized with whole virus vaccine followed by viral proteins, 75 and 56kDa in
combination which was comparable with birds immunized with 56kDa protein alone.
Despite lower values of HI and ELISA titers elicited by viral subunits in
immunized birds, when challenged with virulent NDV strain, protection accorded
by viral proteins in combination (75 +56kDa) or 56kDa alone was comparable with
whole virus vaccine.

Indian Journal of Experimental Biology

Vol. 48, September 2010, pp. 896-904

Antioxidant and hepatoprotective effect of the ethyl acetate
extract of Enicostemma axillare
(Lam). Raynal against CCl4-induced liver injury in rats

Enicostemma
axillare is used in
Indian traditional medicine as a liver tonic. Its ethyl acetate extract has
shown potent in vitro antioxidant activity and found to contain 7.26% of
a bitter secoiridoid glycoside, swertiamarin. Hence, in the present study the
ethyl acetate extract was screened for hepatoprotective and antioxidant
properties against CCl4 induced hepatic injury in rats. The
hepatoprotection was assessed in terms of reduction in histological damage and
changes in serum enzymes and metabolites. The pretreatment with the extract at
100 and 200 mg/kg body weight doses given orally for eight days prior to CCl4
caused significant restoration of altered biochemical changes due to CCl4
towards the normal in serum, liver and kidney. The extract treatment at 200
mg/kg body weight was found to be more potent than the standard silymarin at
100 mg/kg body weight in reversing most of the biochemical parameters.
Histopathological studies complemented the results of biochemical estimations
in providing a proof of hepatoprotective and antioxidant actions of the
extract. The study provides a support to the ethnomedical use of E. axillare
in India.

Aim of the study was to investigate the
methanol and aqueous extracts of heartwood of C. sappan for its
hepatoprotective activity against CCl4 induced toxicity in freshly
isolated rat hepatocytes and animals. Freshly isolated rat hepatocytes were
exposed to CCl4 (1%) along with/without various concentrations of
methanolic and aqueous extract of C. sappan (1000-800 µg/ml) and the levels of selected liver enzymes were
estimated. Antihepatotoxic effect of methanolic extract was observed in freshly
isolated rat hepatocytes at concentrations 1000-800 µg/ml and was found to be similar
to that of standard drug silymarin. Wistar strain albino rat model was used for
the investigation of in vivo hepatoprotective
properties of aqueous and methanolic extract of C. sappan (100 and 200 mg/kg body weight). Liver damage was induced
by ip administration of CCl4 (30%) suspended in olive oil (1 ml/kg
body weight). Both the tested extracts showed potent hepatoprotective activity at
200 mg/kg body weight test dose which was comparable with that of the standard
silymarin used in similar test dose. The methanolic and aqueous extract was
able to restore the biochemical levels to normal which were altered due to CCl4
intoxication in freshly isolated rat hepatocytes and also in animals.

Therapeutic
effect of ethanolic extract of Hygrophila
spinosa in gentamicin-induced nephrotoxic model of kidney injury in male
Sprague-Dawley rats was studied. Rats were administered with gentamicin at a
dose of 80 mg/kg intraperitoneally (ip) to induce nephrotoxicity. Kidney
function was assessed by measuring serum creatinine and urea. Kidney superoxide
dismutase, lipid peroxidation, catalase and reduced glutathione were also
measured in control and treated rats. H. spinosa extract showed free
radical scavenging activities at doses of 50 and 250 mg/kg with a predominant
activity at
250 mg/kg. The ethanolic extract also caused a reduction in serum creatinine
and urea levels. Histopathological studies were conducted to confirm the
therapeutic action of the plant extract. The results demonstrated that the
ethanolic extract of whole plant of H.
spinosa evinced the therapeutic effect and inhibited gentamicin-induced
proximal tubular necrosis.

The petroleum ether (PE), chloroform (CH),
ethanol (ETH) and water extracts of Terminalia
bellerica and T. chebula fruits
were evaluated for their analgesic activity using the tail immersion model in
mice. The ethanolic extracts of both the plants exhibited analgesic response at
200,400 and 800mg/kg. The studies were further carried for 15 days to evaluate
the effect of these extracts in chronic pain and maximum analgesic response was
observed on 14th day in both the plants. Phytochemical investigation
of ethanolic extract of the fruits of Terminalia
bellerica and T. chebula revealed
the presence of saponins, triterpenoids, carbohydrates, tannins and proteins.
The results indicate that fruits of T.
bellerica and T. chebula could be
considered as potential candidate for bioactivity-guided isolation of natural
analgesic agents used in the management of chronic pain.

Indian Journal of Experimental Biology

Vol. 48, September 2010, pp. 931-935

In vitro effect of some Indian honeys on Staphylococcus
aureus from wounds

Staphylococcus
aureus is the most frequently isolated pathogen
from wounds with multiple resistances to antibiotics. Honey has been
demonstrated and reported to be effective antibacterial agent on Gram positive
and Gram negative organisms. Hence, the present study was conducted to evaluate
the in vitro antibacterial effect of
Indian honeys on Staphylococcus aureus
obtained from wounds. A total of 123 Staphylococcus
aureus isolates along with ATCC 25923 were categorized as sensitive, multi
drug resistant (MDR) and non-MDR strains. Out of total nine Indian honeys
(three each of unifloral, multifloral and branded marketed honey) used, three
unifloral and three multifloral honey samples showed antibacterial activity
against all the organisms tested by Agar diffusion method but not the branded
marketed honeys. The MIC values of all honey samples for all studied Staphylococcus aureus isolates ranged
between 5-15% (v/v). Unifloral honey samples showed higher antibacterial
activity than multifloral honey. The single sample of Jambhul honey showed the
highest activity. Thus, Indian honeys were found to be effective for their
antimicrobial activity on sensitive, non-MDR, MDR and ATCC strains of S. aureus.

Indian Journal of Experimental Biology

Vol. 48, September 2010, pp. 936-941

Optimization of medium for lipase production by Acinetobacter
haemolyticus from healthy human skin

3Institute
of Bioinformatics and Biotechnology, University
of Pune 411 007, India.

Received 25 November 2009, revised 18 May 2010

A lipase
producing Acinetobacter haemolyticus
TA106 was isolated from healthy human skin of tribal population. The maximum
activity of 55 U/ml was observed after medium optimization using the “one
variable at a time” and the statistical approaches. The optimal composition of
the medium was determined as (% w/v or v/v): tryptone - 1, yeast extract - 0.5,
sodium chloride-1, olive oil-1, Tween - 80 1, manganese sulphate - 5 mM,
sucrose- 1, pH-7. It was found that
maximum production occurred in late log phase i.e. after 72 h and at 200 rpm.
From factorial design and statistical analysis, it was found that pH, temperature, salt, inoculum density
and aeration significantly affected the lipase production. It was also noted
that inoculum density of 3 % (v/v), sucrose (1% w/v) and manganese sulphate(5 mM) displayed maximum lipase activity of 55
U/ml by conventional as well as statistical method. Optimization studies also
indicated the increase in specific activity from 0.2 U/mg to 6.7 U/mg.

Indian Journal of Experimental Biology

Vol. 48, September 2010, pp. 942-947

Poly-β-hydroxybutyrate production by Pseudomonas sp. RZS 1 under aerobic and
semi-aerobic condition

Pseudomonas sp. RZS1 was isolated from distillery effluent and identified based
on phenotypic characters and 16s rRNA sequencing. It accumulated optimum amount
(703.79 µg/mg of biomass) of poly-β-hydroxybutyrate(PHB)under aerobic
process of fermentation and 75 µg/mg of biomass under the anaerobic process of
fermentation. Aerobic fermentation yielded 9.3-fold more PHB than semi-aerobic
fermentation. Acetone alcohol method proved to be the best suitable recovery
method as it gave 703.79 µg PHB per mg of biomass with a percentage recovery
yield of 70.37. It started to accumulate PHB at the end of lag phase (from 6 h
of incubation). Optimum amount of PHB (20 µg/ml) was reported during early
stationary phase (30 h of incubation). Extracted PHB showed two peaks, minor
one at 248 nm and major one at 365 nm. IR spectra revealed the presence of
functional groups characteristics of PHB.

Indian Journal of Experimental Biology

Vol. 48, September 2010, pp. 948-950

Notes

Comparative evaluation of PCR
in Ziehl-Neelsen stained smears and PCR in tissues for diagnosis of Mycobacterium avium subsp. paratuberculosis

4Institute ofBiotechnology
and Bioengineering, Centre of Genetics and
Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD), 5001-801 Vila Real, Portugal

Received
3 December 2009; revised 8 April 2010

Thirty six tissues from
sheep, previously diagnosed with paratuberculosis, were tested by PCR in positive Ziehl-Neelsen
staining smears of tissues, and PCR in tissues targeting IS900 specific for Mycobacterium avium subsp. paratuberculosis. DNA amplification was achieved in 33.3%
Ziehl-Neelsen smears, and in 61.1% tissue samples. Combination of both techniques found 66.7%
samples as positive. Combination of techniques would, therefore, increase the
sensitivity of diagnosis.